【目的】对限制性位点扩增多态性（Restriction site amplified polymorphism，RsAP）标记技术进行改进，并优化了其反应体系。【方法】遵循引物设计原则，使限制性位点序列位于中间，在其3’端增加3个选择性碱基，5’端设计为10～12个...【目的】对限制性位点扩增多态性（Restriction site amplified polymorphism，RsAP）标记技术进行改进，并优化了其反应体系。【方法】遵循引物设计原则，使限制性位点序列位于中间，在其3’端增加3个选择性碱基，5’端设计为10～12个碱基的随机序列，设计了14条长为19bp的限制性位点扩增多态性（RSAP）新引物。以大白菜、紫菜薹自交系及其F1、F2的DNA为模板，对新引物的反应体系进行了优化，并对其重复性和适用性进行了检验。【结果】新设计引物PCR扩增的前5个循环退火温度为35℃，后35个循环为52℃；在25μL优化反应体系中，模板DNA用量为2．0μL（20．0ng／μL）、Mg^3＋ 3．0μL（25mmol／L）、Taq酶（5U／μL）1．5U、dNTPs2．0μL（2．5mmol／L）、引物各0．6μL（10μmol／L）；新引物扩增出的谱带较原引物更多，多态性更好；只改变选择性碱基，新设计引物就能扩增出新的谱带；新设计引物的重复性、稳定性好，在荞麦和小麦品系，以及紫菜薹、大白菜及其F1、F2中，新引物均能扩增出清晰谱带。【结论】新引物适用性和重复性好，可广泛应用于其他作物的基因组DNA分析。展开更多
Stoichiometry has long been addressed in the studies of ecosystem ecology, but it was almost ignored for a long time. Until recently, ecologists have become aware that stoichiometry could provide a new tool to study e...Stoichiometry has long been addressed in the studies of ecosystem ecology, but it was almost ignored for a long time. Until recently, ecologists have become aware that stoichiometry could provide a new tool to study ecology from genes to the biosphere. Among this trend, N:P stoichiometry is used actively in ecological interactions since nitrogen (N) and phosphorus (P) are the two most important elements in most ecosystems. This article reviews the application of N:P stoichiometry to the studies of ecological problems at different levels, including ecosystem, community and species. Meanwhile, we also provide the cellular basis of N:P stoichiometry, identify the shortages in the use of N:P stoichiometry theory, and put forward some perspectives for future research to be conducted.展开更多
利用8个代表性菌株从153对引物组合中筛选出多态性高且稳定性好的10对引物组合,对来自黑龙江省各水稻主要产区的214个稻瘟病菌菌株进行RSAP(Restriction site amplified polymorphism)分析,得到134条多态性条带,经聚丙烯酰胺凝胶电泳,得...利用8个代表性菌株从153对引物组合中筛选出多态性高且稳定性好的10对引物组合,对来自黑龙江省各水稻主要产区的214个稻瘟病菌菌株进行RSAP(Restriction site amplified polymorphism)分析,得到134条多态性条带,经聚丙烯酰胺凝胶电泳,得到196个不同单元型(带型)的代表性菌株的指纹图谱。用UPGMA进行聚类分析,以相似系数0.83为界,将214个菌株196个单元型划分为10个遗传宗谱,10个遗传宗谱在黑龙江省的分布存在明显地域差异。展开更多
The variation of microbial community structure was investigated for the tank bioleaching process of Pb-Zn-Sn chalcopyrite concentrate in the presence of mixed moderately thermophilic bacteria. The parameters, such as ...The variation of microbial community structure was investigated for the tank bioleaching process of Pb-Zn-Sn chalcopyrite concentrate in the presence of mixed moderately thermophilic bacteria. The parameters, such as pH value, solution potential and concentrations of metal ions, were determined by the method of polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） to analyze the succession of microbial community. The results showed that a final copper extraction rate of 85.6% could be obtained after tank bioleaching for 30 d. The Acidithiobacillus caldus was the dominant population with abundance of about 73.80%in the initial stage, then Sulfobacillus thermosulfidooxidans dominated from the 18th day to the end of bioleaching, while the abundance of Leptospirillum ferriphilum changed slightly. A higher solution potential within a certain range and appropriate concentration of ferric ions were essential for this tank bioleaching of chalcopyrite.展开更多
AIM: To study the polymorphisms of toll-like receptor 4 (TLR4) gene Asp299Gly, Thr399Ile and TLR2 gene Arg753Gln, Arg677Trp and susceptibility to inflammatory bowel disease (IBD) in the Zhuang population from Guangxi,...AIM: To study the polymorphisms of toll-like receptor 4 (TLR4) gene Asp299Gly, Thr399Ile and TLR2 gene Arg753Gln, Arg677Trp and susceptibility to inflammatory bowel disease (IBD) in the Zhuang population from Guangxi, China. METHODS: A case-control study was performed from February 2007 to October 2011 which included 146 Zhuang patients with IBD in the experimental group and 164 healthy Zhuang subjects who acted as the control group. All patients and healthy subjects were from the Guangxi Zhuang Autonomous Region of China. Genomic DNA was extracted from intestinal tissue by the phenol chloroform method. TLR4 gene Asp299Gly, Thr399Ile and TLR2 gene Arg753Gln, Arg677Trp were amplified by polymerase chain reaction (PCR), and then detected by PCR-restriction fragment length polymorphism (RFLP). RESULTS: The TLR4 gene Asp299Gly was digested using Nco Ⅰ restriction enzyme, and a single band of 249 bp was observed which showed that it was a wild type (AA). The TLR4 gene Thr399Ile was digested using Hinf Ⅰrestriction enzyme and only the wild type (CC) was detected. In addition, the TLR2 gene Arg-677Trp was digested using Aci Ⅰ restriction enzyme and only the wild type (CC) was detected. The TLR2 gene Arg753Gln was digested using Pst Ⅰ restriction enzyme. Only the wild type (GG) as a single band of 254 bp was observed during RFLP. Overall, no heterozygous or homozygous single nucleotide polymorphism mutations were found in patients with Crohn's disease and ulcerative colitis both in the TLR4 gene Asp299Gly, Thr399Ile and the TLR2 gene Arg677Trp, Arg753Gln in the Zhuang population from the Guangxi Zhuang Autonomous Region of China. CONCLUSION: The TLR4 gene Asp299Gly, Thr399Ile and TLR2 gene Arg753Gln, Arg677Trp polymorphisms may not be associated with IBD in the Zhuang population from the Guangxi Zhuang Autonomous Region of China.展开更多
文摘【目的】对限制性位点扩增多态性（Restriction site amplified polymorphism，RsAP）标记技术进行改进，并优化了其反应体系。【方法】遵循引物设计原则，使限制性位点序列位于中间，在其3’端增加3个选择性碱基，5’端设计为10～12个碱基的随机序列，设计了14条长为19bp的限制性位点扩增多态性（RSAP）新引物。以大白菜、紫菜薹自交系及其F1、F2的DNA为模板，对新引物的反应体系进行了优化，并对其重复性和适用性进行了检验。【结果】新设计引物PCR扩增的前5个循环退火温度为35℃，后35个循环为52℃；在25μL优化反应体系中，模板DNA用量为2．0μL（20．0ng／μL）、Mg^3＋ 3．0μL（25mmol／L）、Taq酶（5U／μL）1．5U、dNTPs2．0μL（2．5mmol／L）、引物各0．6μL（10μmol／L）；新引物扩增出的谱带较原引物更多，多态性更好；只改变选择性碱基，新设计引物就能扩增出新的谱带；新设计引物的重复性、稳定性好，在荞麦和小麦品系，以及紫菜薹、大白菜及其F1、F2中，新引物均能扩增出清晰谱带。【结论】新引物适用性和重复性好，可广泛应用于其他作物的基因组DNA分析。
文摘Stoichiometry has long been addressed in the studies of ecosystem ecology, but it was almost ignored for a long time. Until recently, ecologists have become aware that stoichiometry could provide a new tool to study ecology from genes to the biosphere. Among this trend, N:P stoichiometry is used actively in ecological interactions since nitrogen (N) and phosphorus (P) are the two most important elements in most ecosystems. This article reviews the application of N:P stoichiometry to the studies of ecological problems at different levels, including ecosystem, community and species. Meanwhile, we also provide the cellular basis of N:P stoichiometry, identify the shortages in the use of N:P stoichiometry theory, and put forward some perspectives for future research to be conducted.
文摘利用8个代表性菌株从153对引物组合中筛选出多态性高且稳定性好的10对引物组合,对来自黑龙江省各水稻主要产区的214个稻瘟病菌菌株进行RSAP(Restriction site amplified polymorphism)分析,得到134条多态性条带,经聚丙烯酰胺凝胶电泳,得到196个不同单元型(带型)的代表性菌株的指纹图谱。用UPGMA进行聚类分析,以相似系数0.83为界,将214个菌株196个单元型划分为10个遗传宗谱,10个遗传宗谱在黑龙江省的分布存在明显地域差异。
基金Project (51374248) supported by the National Natural Science Foundation of ChinaProject (NCET-13-0595) supported by the Program for New Century Excellent Talents in University,China+3 种基金Project (2012AA061501) supported by the High-tech Research and Development Program of ChinaProject (2010CB630905) supported by the National Basic Research Program of ChinaProject (20120162120010) supported by the Research Fund for the Doctoral Program of Higher Education of ChinaProject (CSUZC2012020) supported by the Open-End Fund for the Valuable in Central South University,China
文摘The variation of microbial community structure was investigated for the tank bioleaching process of Pb-Zn-Sn chalcopyrite concentrate in the presence of mixed moderately thermophilic bacteria. The parameters, such as pH value, solution potential and concentrations of metal ions, were determined by the method of polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） to analyze the succession of microbial community. The results showed that a final copper extraction rate of 85.6% could be obtained after tank bioleaching for 30 d. The Acidithiobacillus caldus was the dominant population with abundance of about 73.80%in the initial stage, then Sulfobacillus thermosulfidooxidans dominated from the 18th day to the end of bioleaching, while the abundance of Leptospirillum ferriphilum changed slightly. A higher solution potential within a certain range and appropriate concentration of ferric ions were essential for this tank bioleaching of chalcopyrite.
基金Supported by The Natural Science Foundation of Guangxi Zhuang Autonomous Region, China, No. 0832009the Chinese Traditional Medicine Science Foundation of Guangxi Zhuang Autonomous Region, China, No. GZKZ10-108
文摘AIM: To study the polymorphisms of toll-like receptor 4 (TLR4) gene Asp299Gly, Thr399Ile and TLR2 gene Arg753Gln, Arg677Trp and susceptibility to inflammatory bowel disease (IBD) in the Zhuang population from Guangxi, China. METHODS: A case-control study was performed from February 2007 to October 2011 which included 146 Zhuang patients with IBD in the experimental group and 164 healthy Zhuang subjects who acted as the control group. All patients and healthy subjects were from the Guangxi Zhuang Autonomous Region of China. Genomic DNA was extracted from intestinal tissue by the phenol chloroform method. TLR4 gene Asp299Gly, Thr399Ile and TLR2 gene Arg753Gln, Arg677Trp were amplified by polymerase chain reaction (PCR), and then detected by PCR-restriction fragment length polymorphism (RFLP). RESULTS: The TLR4 gene Asp299Gly was digested using Nco Ⅰ restriction enzyme, and a single band of 249 bp was observed which showed that it was a wild type (AA). The TLR4 gene Thr399Ile was digested using Hinf Ⅰrestriction enzyme and only the wild type (CC) was detected. In addition, the TLR2 gene Arg-677Trp was digested using Aci Ⅰ restriction enzyme and only the wild type (CC) was detected. The TLR2 gene Arg753Gln was digested using Pst Ⅰ restriction enzyme. Only the wild type (GG) as a single band of 254 bp was observed during RFLP. Overall, no heterozygous or homozygous single nucleotide polymorphism mutations were found in patients with Crohn's disease and ulcerative colitis both in the TLR4 gene Asp299Gly, Thr399Ile and the TLR2 gene Arg677Trp, Arg753Gln in the Zhuang population from the Guangxi Zhuang Autonomous Region of China. CONCLUSION: The TLR4 gene Asp299Gly, Thr399Ile and TLR2 gene Arg753Gln, Arg677Trp polymorphisms may not be associated with IBD in the Zhuang population from the Guangxi Zhuang Autonomous Region of China.
